The nuclear receptor superfamily : methods and protocols / edited by Iain J. McEwan. — New York : Humana Press, c2009. – (58.17/M592/v.505)

Contents

    Contents
    
    Preface v
    Contributors ix
    PART I: INTRODUCTION
    1 Nuclear Receptors: One Big Family 3
    PART II: LIGAND BINDING AND NUCLEAR RECEPTOR TUKNOWR
    2 Methods for Measuring Ligand Dissociation and Nuclear Receptor Turnover in Whole Cells 21
    3 Flow Cytometry as a Tool for Measurement of Steroid Hormone Receptor Protein Expression in Leukocytes 35
    4 X-Ray Crystallography of Agonist/Antagonist-Bound Receptors 51
    PART III: NUCLEAR LOCALIZATION AND DNA BINDING
    5 FRAP and FRET Methods to Study Nuclear Receptors in Living Cells 69
    6 Receptor-DNA Interactions: EMSA and Footprinting 97
    7 Chromatin Immunoprecipitation (CHIP) Methodology and Readouts 123
    PART IV: NUCLEAR RECEPTOR -- CO-REGULATORY PROTEIN INTERACTIONS
    8 Yeast-Based Reporter Assays for the Functional Characterization of Cochaperone Interactions with Steroid Hormone Receptors 141
    9 High Throughput Analysis of Nuclear Receptor-Cofactor Interactions 157
    10 Binding Affinity and Kinetic Analysis of Nuclear Receptor/Co-Regulator Interactions Using Surface Plasmon Resonance 171
    11 Using RNA Interference to Study Protein Function
    12 Using Intrinsic Fluorescence Emission Spectroscopy to Study Steroid Receptor and Coactivator Protein Conformation Dynamics 205
    PART V: PATHOPHYSIOLOGICAL ANALYSIS OF NUCLEAR RECEPTOR FUNCTION
    13 Development of Phosphorylation Site-Specific Antibodies to Nuclear Receptors
    14 Tissue-Selective Knockouts of Steroid Receptors: A Novel Paradigm in the Study of Steroid Action
    15 Methods for Identifying and Studying Genetic Alterations in Hormone-Dependent Cancers